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目的 气管干细胞的原位观察。 方法 使用 5 氟尿嘧啶 (5 FU)造成大鼠离体气管环的严重损伤 ,采用光镜、PCNA免疫组织化学 ,以及Hoechst33342染色观察气管黏膜的修复过程。 结果 5 FU作用 12h后 ,气管上皮脱落 ,可见少量间隔分布的类似裸核的细胞呈钉状位于基底膜上 ,PCNA染色阴性 (G0 期细胞 ) ,其中少数细胞Hoechst33342染色阴性。去除 5 FU 6h后 ,气管黏膜由扁平上皮覆盖 ;4 8h后 ,气管环恢复假复层纤毛柱状上皮。结论 5 FU能杀死处于细胞周期的气管上皮细胞 ,对G0 期细胞作用很小 ,残留于基底膜上的裸核样的G0 期细胞中含有干细胞 ,其Hoechst33342染色阴性 ,并具有排出荧光染料的能力 ,正是这些细胞的增殖分化 ,修复了损伤的气管环。
Objective To observe the tracheal stem cells in situ. Methods The rat tracheal rings were severely injured by 5-fluorouracil (5 FU). The repair process of tracheal mucosa was observed by light microscopy, PCNA immunohistochemistry and Hoechst33342 staining. Results After 12 hours of 5 FU tracheal epithelial detachment, a small number of naked nucleus-like cells with spikes were located on the basement membrane. PCNA staining was negative (G0 stage cells), and a few cells were negative for Hoechst33342 staining. Tracheal mucosa was covered by flat epithelium after removing 5 FU for 6 hours. After 48 hours, the tracheal ring restored pseudostratified ciliated columnar epithelium. Conclusions 5 FU can kill the tracheal epithelial cells in the cell cycle and have little effect on the cells in the G0 phase. The naked nucleated G0 cells remaining on the basement membrane contain stem cells with negative Hoechst33342 staining and fluorescein-emitting Capacity, it is these cells proliferation and differentiation, repair damaged tracheal ring.